• Julio Celis, Danish Cancer Society; Institute of Cancer Biology and Danish Centre for Human Genome Research; Copenhagen, Denmark

Protein assay methods are used for protein identification with blood groups, cell surface markers, drugs and toxins. This text features comprehensive protocols essential for researchers studying various areas of biological and medical sciences. The techniques in this text are presented in a friendly step-by-step fashion, providing useful tips and potential pitfalls while enabling researchers at all stages to embark on basic problems using a vareity of technologies and model systems.
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Graduate students and professionals in a lab enviornment


Book information

  • Published: November 2009
  • ISBN: 978-0-12-375692-3

Table of Contents

  1. Protein Determination
  2. Phosphopeptide Mapping: A Basic Protocol
  3. Coupling of Fluorescent Tags to Proteins
  4. Radioiodination of Proteins and Peptides
  5. Free-Flow Electrophoresis
  6. Gel-Based Proteomics: High-Resolution Two-Dimensional Gel Electrophoresis of Proteins. Isoelectric Focusing and Nonequalibrium pH Gradient Electrophoresis.
  7. High-Resolution Two-Dimensional Electrophoresis with Immobilized pH Gradients for Proteome Analysis
  8. Two-Dimensional Difference Gel Electrophoresis: Application for the Analysis of Differential Protein Expression in Multiple Biological Samples
  9. Affinity Electrophoresis for Studies of Biospecific Interations: High-Resolution Two-Dimensional Affinity Electrophoresis for Separation of Hapten-Specific Polyclonal Antibodies into Monoclonal Antibodies in Murine Blood Plasma
  10. Image Analysis and Quantitation
  11. Fluorescence Detection of Proteins in Gels Using SYPRO Dyes
  12. Autoradiography and Fluorography: Film-Based Techniques for Imaging Radioactivity in Flat Samples
  13. Two-Dimensional Gel Profiling of Posttanslationally Modified Proteins by in vivo Isotope Labeling
  14. Immunoprecipitation of Proteins under Nondenaturing Conditions
  15. Nondenaturing Polyacrylamide Gel Electrophoresis as a Method for Studying Protein Interations: Applications in the Analysis of Mitochrondrial Oxidative Phosphorylation Complexes
  16. Affinity purification with Natural Immobilized Ligands
  17. Analysis of Protein-Protein Interactions by Chemical Cross-Linking
  18. Peroxisomal Targeting as a Tool to Assess Protein-Protein Interactions
  19. Biomolecular Interaction Analysis Mass Spectrometry
  20. Blot Overlays with P-Labedled GST-Ras Fusion Proteins: Application to Mapping Protein-Protein Interaction Sites
  21. Ligand Blot Overlay Assay: Detetion of Ca+2- and Small GTP-Binding Proteins
  22. Modular Scale Yeast Two-Hybrid Screening
  23. Chromophore-Assisted Laser Inactivation of Proteins by Antibodies Labeled with Malachite Green
  24. Chromatin Immunoprecipitation (ChIP)
  25. Gel Mobility Shift Assay
  26. DNA Affinity Chromatography of Transcription Factors: The Oligaonucleotide Trapping Approach
  27. Protein Degradation Methods: Chaperone-Mediated Autophagy
  28. Methods in Protein Ubiquitination
  29. Protein Identification and Sequencing by Mass Spectrometry
  30. Proteome Specific Sample Preparation Methods for Matrix-Assisted Laser Desorption/Ionization Mass Spectrometry
  31. In-Gel Digestion of Protein Spots for Mass Spectrometry
  32. Peptide Sequencing by Tandem Mass Spectrometry
  33. Direct Database Searing Using Tandem Mass Spectra of Peptides
  34. Identification of Proteins from Organisms with Unsequenced Genomes by Tandem Mass Spectrometry and Sequence-Similarity Database Searching Tools
  35. Identification of Protein Phosphorylation Sites by Mass Spectrometry
  36. Analysis of Carbohydrates/Glycoproteins by Mass Spectrometry
  37. Stable Isotope Labeling by Amindo Acids in Cell Culture for Quantitative Proteomics
  38. Site-Specific, Stable Isotopic Labeling of Cysteinyl Pepties in Complex Peptide Mixtures
  39. Protein Hydrogen Exchange measured by Electrospry Ionization Mass Spectrometry
  40. Nongel Based Proteomics: Selective Reversed-Phase Chromatographic Isolation of Methionine-Containing Peptides from Complex Peptide Mixtures
  41. Mass Spectrometry in Noncovalent Protein Interactions and Protein Assemblies